[HLA-DR typing using PCR-SSO with simultaneous hybridization of each dot blot membrane with a digoxigenin- and a biotin-marked probe].
Here we present a PCR-SSO procedure designed to simplify HLA-DR typing. We labelled 8 of a total of 16 oligonucleotide probes with digoxigenin, the others with biotin, and formed 8 pairs, each containing a digoxigenin- as well as a biotin-labelled probe. Each pair was hybridized simultaneously to one of eight dot blot membranes containing the DNA to be typed. Specific binding was detected by incubation with a mixture of the appropriate conjugates followed by sequential addition first of a chemiluminescent substrate to detect the digoxigenin-labelled probe and then a chromogenic substrate for detection of the biotin-labelled probe. With this procedure, the specific binding of two different probes to the same dot blot membrane could be evaluated, considerably reducing the labor inherent in PCR-SSO typing.
Chen
D F
DF
Blutbank-Immunhämatologie-Transfusionsmedizin, Medizinische Hochschule Hannover, Deutschland.
Endres
W
W
Meyer
S A
SA
Stangel
W
W
D001710:Biotin; D004076:Digoxigenin; D006684:HLA-DR Antigens / Q000235:genetics*; D006650:Histocompatibility Testing / Q000379:methods*; D006801:Humans; D017403:In Situ Hybridization / Q000379:methods; D015345:Oligonucleotide Probes; D016133:Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity
D006684:HLA-DR Antigens; D015345:Oligonucleotide Probes; D001710:Biotin; D004076:Digoxigenin
HLA-DR-Typisierung mittels PCR-SSO bei gleichzeitiger Hybridisierung jeder Dot-blot-Membran mit einer Digoxigenin- und einer Biotin-markierten Sonde.